Skip to main content

Apricot (Prunus armeniaca L.)

  • Protocol
  • First Online:
Agrobacterium Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1224))

Abstract

A protocol for Agrobacterium-mediated stable transformation of whole leaf explants of the apricot (Prunus armeniaca) cultivars ‘Helena’ and ‘Canino’ is described. Regenerated buds were selected using a two-step selection strategy with paromomycin sulfate and transferred to bud multiplication medium 1 week after they were detected for optimal survival. After buds were transferred to bud multiplication medium, antibiotic was changed to kanamycin and concentration increased gradually at each transfer to fresh medium in order to eliminate possible escapes and chimeras. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines, was 5.6 %. Green and healthy buds, surviving high kanamycin concentration, were transferred to shoot multiplication medium where they elongated in shoots and proliferated. Elongated transgenic shoots were rooted in a medium containing 70 μM kanamycin. Rooted plants were acclimatized following standard procedures. This constitutes the only transformation protocol described for apricot clonal tissues and one of the few of Prunus.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Petri C, Burgos L (2005) Transformation of fruit trees. Useful breeding tool or continued future prospect? Transgenic Res 14:15–26

    Article  CAS  PubMed  Google Scholar 

  2. Pérez-Tornero O, Burgos L (2000) Different media requirements for micropropagation of apricot cultivars. Plant Cell Tiss Organ Cult 63:133–141

    Article  Google Scholar 

  3. Pérez-Tornero O, Burgos L, Egea J (1999) Introduction and establishment of apricot in vitro through the regeneration of shoots from meristem tips. In Vitro Cell Dev Biol Plant 35:249–253

    Article  Google Scholar 

  4. Pérez-Tornero O, Egea J, Vanoostende A, Burgos L (2000) Assessment of factors affecting adventitious shoot regeneration from in vitro cultured leaves of apricot. Plant Sci 158:61–70

    Article  PubMed  Google Scholar 

  5. Burgos L, Alburquerque N (2003) Low kanamycin concentration and ethylene inhibitors improve adventitious regeneration from apricot leaves. Plant Cell Rep 21:1167–1174

    Article  CAS  PubMed  Google Scholar 

  6. Petri C, Alburquerque N, García-Castillo S, Egea J, Burgos L (2004) Factors affecting gene transfer efficiency to apricot leaves during early Agrobacterium-mediated transformation steps. J Hortic Sci Biotechnol 79:704–712

    CAS  Google Scholar 

  7. Petri C, Alburquerque N, Pérez-Tornero O, Burgos L (2005) Auxin pulses and a synergistic interaction between polyamines and ethylene inhibitors improve adventitious regeneration from apricot leaves and Agrobacterium-mediated transformation of leaf tissues. Plant Cell Tiss Organ Cult 82:105–111

    Article  CAS  Google Scholar 

  8. Petri C, Alburquerque N, Burgos L (2005) The effect of aminoglycoside antibiotics on the adventitious regeneration from apricot leaves and selection of nptII-transformed leaf tissues. Plant Cell Tiss Organ Cult 80:271–276

    Article  CAS  Google Scholar 

  9. Petri C, Wang H, Alburquerque N, Faize M, Burgos L (2008) Agrobacterium-mediated transformation of apricot (Prunus armeniaca L.) leaf explants. Plant Cell Rep 27:1317–1324

    Article  CAS  PubMed  Google Scholar 

  10. Petri C, López-Noguera S, Alburquerque N, Egea J, Burgos L (2008) An antibiotic-based selection strategy to regenerate transformed plants from apricot leaves with high efficiency. Plant Sci 175:777–783

    Article  CAS  Google Scholar 

  11. Petri C et al (2012) A chemical-inducible Cre-LoxP system allows for elimination of selection marker genes in transgenic apricot. Plant Cell Tiss Organ Cult 110:337–346

    Article  CAS  Google Scholar 

  12. López-Noguera S, Petri C, Burgos L (2009) Combining a regeneration-promoting gene and site-specific recombination allows a more efficient apricot transformation and the elimination of marker genes. Plant Cell Rep 28:1781–1790

    Article  PubMed  Google Scholar 

  13. Chiu C et al (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330

    Article  CAS  PubMed  Google Scholar 

  14. Vancanneyt G, Schmidt R, O’Connor-Sanchez A, Willmitzer L, Rocha-Sosa M (1990) Construction of an intron-containing marker gene: Splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant transformation. Mol Gen Genet 220:245–250

    Article  CAS  PubMed  Google Scholar 

  15. Quoirin M, Lepoivre P (1977) Etude de milieux adaptes aux cultures in vitro de Prunus. Acta Hortic 78:437–442

    Google Scholar 

  16. Driver JA, Kuniyuki AH (1984) In vitro propagation of Paradox walnut rootstock. HortScience 19:507–509

    Google Scholar 

  17. Alt-Mörbe J, Kühlmann H, Schröder J (1989) Differences in induction of Ti plasmid virulence genes virG and virD and continued control of virD expression by four external factors. Mol Plant-Microbe Interact 2:301–308

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lorenzo Burgos .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Petri, C., Alburquerque, N., Burgos, L. (2015). Apricot (Prunus armeniaca L.). In: Wang, K. (eds) Agrobacterium Protocols. Methods in Molecular Biology, vol 1224. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-1658-0_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1658-0_10

  • Published:

  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-1657-3

  • Online ISBN: 978-1-4939-1658-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics